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Research ArticleOpen Access

Extraction of Inonotus obliquus strains Genomic DNA and Optimization of RAPD System

Volume 2 - Issue 1

Guo Xiaofan1 and Wang Shouming*2

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    • 1College of Life Engineering, Shenyang Institute of Technology, China
    • 2College of Horticulture, Shenyang Agricultural University, China

    *Corresponding author: Wang Shouming, College of Horticulture, Shenyang Agricultural University, Shenyang 110866, China, Email: smwang0206@163.com

Received: January 04, 2018;   Published: January 12, 2018

DOI: 10.26717/BJSTR.2018.02.000657

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Abstract

The strains of wild Inonotus obliquus were used as the materials. An improved CTAB method of Inonotus obliquus DNA extraction was applied to obtaining genomic DNA. And it was used for template then to optimize RAPD amplification conditions. The results indicated that: a volume of 25 μL was used which contained 40ng/μl template DNA, 10pmol primer, 2mmol/L Mg2+, 1.0 unit domestic Taq DNA polymerase, 100μmol/L dNTP, others complement with ddH2O. The amplification program was that 1 cycle of 5 minutes at 94oC to initial denature; 45 cycles of 1 minute at 94oC to denature, 1 minute at 40oC for annealing, 1.5 minutes at 72oC for extension; 5 minutes at 72oC for final extension.

Keywords: Amplification; Fungus; Mycelium; Primer Screening

Abbreviations: RAPD: Random Amplification of Polymorphic DNA; PDA: Potato Dextrose Agar

Abstract| Introduction| Materials and Methods| Result and Analysis| Discussion| Acknowledgment| References|